LOCUS       Exported                5131 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  In vivo visualization of mitochondria (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    4xmts-mNeonGreen
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5131)
  AUTHORS   Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, 
            Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J
  TITLE     Robust and Bright Genetically Encoded Fluorescent Markers for 
            Highlighting Structures and Compartments in Mammalian Cells
  JOURNAL   bioRxiv 160374
REFERENCE   2  (bases 1 to 5131)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5131
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      122..131
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             complement(198..272)
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             complement(294..368)
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             complement(396..470)
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             complement(492..566)
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     primer_bind     complement(579..598)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(580..598)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(643..846)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(672..692)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        847..1150
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1325..1913)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1405..1424)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2242..2289
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2314..2333
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2521..3315)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2632..2651)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3242..3261
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3350..3707)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3364..3499
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3418..3437)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3664..3684
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(3709..3813)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3840..4295
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3927..3946)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4137..4158
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4302..4423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4313..4332)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4367..4386
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4546..125)
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      4992..5001
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 tcatttggat tgccggtgcc ctgacccacc atgtcaaagt ccacaccgtt gatggagcca
       61 aagatgtgta actcatgtgt cgctgggaga gaggccatgt tatcctcctc gcccttgctc
      121 accatggtgg cgaccggtgg atcccgggcc cgcggtaccg tcgactgcag aattcgaagc
      181 ttccccaacg aatggatctt ggcgcgcggc actgggagcc gccgggccga gcctgtcaag
      241 ccccgcagca gcagcggcgt caggacggac atgggatccc ccaacgaatg gatcttggcg
      301 cgcggcactg ggagccgccg ggccgagcct gtcaagcccc gcagcagcag cggcgtcagg
      361 acggacatgg tggcaagctt ccccaacgaa tggatcttgg cgcgcggcac tgggagccgc
      421 cgggccgagc ctgtcaagcc ccgcagcagc agcggcgtca ggacggacat gggatccccc
      481 aacgaatgga tcttggcgcg cggcactggg agccgccggg ccgagcctgt caagccccgc
      541 agcagcagcg gcgtcaggac ggacataagc ttgggtctcc ctatagtgag tcgtattaat
      601 ttcgataagc cagtaagcag tgggttctct agttagccag agagctctgc ttatatagac
      661 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
      721 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
      781 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
      841 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
      901 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
      961 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     1021 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     1081 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     1141 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     1201 ttaataacta atgcatggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1261 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1321 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1381 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1441 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1501 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1561 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1621 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1681 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1741 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1801 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1861 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1921 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1981 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2041 ttaaatcaat ctaaagtata tatgagtaac ctgaggctat ggcagggcct gccgccccga
     2101 cgttggctgc gagccctggg ccttcacccg aacttggggg gtggggtggg gaaaaggaag
     2161 aaacgcgggc gtattggccc caatggggtc tcggtggggt atcgacagag tgccagccct
     2221 gggaccgaac cccgcgttta tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt
     2281 ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc
     2341 tccccctagg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     2401 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     2461 aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc
     2521 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     2581 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     2641 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     2701 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
     2761 gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc
     2821 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     2881 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     2941 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     3001 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     3061 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     3121 cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     3181 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     3241 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     3301 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcgatct ttgcaaaagc
     3361 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     3421 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     3481 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     3541 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     3601 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     3661 cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcaggac tcttcctttt
     3721 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3781 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     3841 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3901 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3961 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4021 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4081 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4141 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4201 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4261 cgcgaatttt aacaaaatat taacgcttac aatttacgcc ttaagataca ttgatgagtt
     4321 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     4381 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     4441 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     4501 ctacaaatgt ggtatggctg attatgatct agagtcgcgg ccgctttact tgtacagctc
     4561 gtccatgccc atcacatcgg taaaggcctt ttgccactcc ttgaagttga gctcggtctt
     4621 ggagtgcttg agctccgtct tacggaacac gtacatcggc tggttcttca gatagttagc
     4681 cgccattggc ttggcaaagg tgtaggtggt ccgcgcagtg ctccggtagc gcttgccatt
     4741 tccagtggtg taactccact taaaggtact gatgatggtt ttgtcgttgg ggtaagtctt
     4801 cttcgacctg caccagtccg cagcggtcag cgagttggtc atcacaggac cgtcagcagg
     4861 gaaaccagtc cccttcacct gggcctctcc tttgatgtgg cttccctcgt aggtgtagcg
     4921 gtagttaaca gtaagggagg caccatcttc aaactgcatt gtgcgatgga cttggtagcc
     4981 ggagccatct accatggcgg cctggaaagg cgacatcccg tcagggtagg gcaggtactg
     5041 atggaagcca tacccgatat gagggaccag aatccagggg gagaactgga ggtcaccctt
     5101 ggtggacttc aggtttaact cctcataacc a
//
